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. 2020 Jul 8;40(28):5402–5412. doi: 10.1523/JNEUROSCI.3190-18.2020

Figure 2.

Figure 2.

Inhibition of CS neuronal activity during postnatal development leads to defects in elimination of vlCST. A, Schematic of a coronal section of the brain showing the injection of AAV8-DREADDs-mCherry into the motor cortex of P2 mice. B, Experimental design. AAV8-DREADDs-mCherry was injected into the motor cortex of P2 mice. Intraperitoneal injection of CNO or saline (as a control) was performed daily from P4 to P14. Brains were analyzed 10 h after intraperitoneal injection at P14. C-J, Representative images of coronal sections from the motor cortex showing immunostaining using antibodies targeting c-fos (green), mCherry (red for hM3Dq), and Ctip2 (blue) from saline- (C-F) or CNO-injected (O-J) hM3Dq-expressing mice at P14. K, Quantification showing the average number of c-fos+ neurons in CNO-injected, hM3Dq-expressing mice is significantly higher than in saline-injected mice (saline group, 44 ± 11.37; CNO group, 112 ± 3.58; p = 0.0013, unpaired t test, n = 4 mice in each group). L-S, Representative images of coronal sections from the motor cortex showing immunostaining using antibodies targeting c-fos (green), mCherry (red for hM4Di), and Ctip2 (blue) from saline- (L-O) or CNO-injected (P-S) hM4Di-expressing mice at P14. T, Quantification showing the average number of c-fos+ neurons in CNO-injected, hM4Di-expressing mice is significantly lower than in saline-injected mice (saline group, 42.75 ± 8.45; CNO group, 9 ± 4.97; p = 0.0013, unpaired t test, n = 4 mice in each group). U, V, Representative images of cervical spinal cords showing the absence of vlCST in the saline-injected (n = 7 mice) (U) group and the presence of vlCST (indicated by red arrows) in the CNO-injected (n = 9 mice) (V) group. W, Quantification of the average area occupied by vlCST in the ipsilateral ventral and lateral funiculi of CNO-injected hM4Di mice shows a significantly larger area than that in saline-injected hM4Di mice (saline group, 0.09999 ± 0.01633%; CNO group, 1.455 ± 0.1971%; p = 0.0003, unpaired t test). Scale bars: J, 50 μm; S, 50 μm; U, 200 μm.